Identification of PKD2 mutations in human preimplantation embryos in vitro using a combination of targeted next-generation sequencing and targeted haplotyping

Chen, Song-Chang; Xu, Xiaoli; Zhang, Junyu; Ding, Guo‐Lian; Jin, Li; Liu, Bei; Sun, Donglin; Mei, Changlin; Yang, Xiaonan; Huang, He‐Feng; Xu, Chenming

doi:10.1038/srep25488

Cited by 15 publications

(8 citation statements)

References 12 publications

(13 reference statements)

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“…Benefited from the WES technology, a lot more pathogenic genes have been found and many types of diseases have been identified111213. Innovative application of new technologies is one of the major factors driving advances in medical science, most clinical applications of next-generation sequencing (NGS) concentrate on known and potential candidate genes to generate clear reports and finally promote clinical diagnosis1415161718. Targeted gene capture and massively parallel sequencing (MPS) have been shown to be an effective technique for genetic analysis and have already led to many exciting discoveries1920.…”

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confidence: 99%

The use of targeted genomic capture and massively parallel sequencing in diagnosis of Chinese Leukoencephalopathies

Wang

Yin

et al. 2016

Sci Rep

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Leukoencephalopathies are diseases with high clinical heterogeneity. In clinical work, it’s difficult for doctors to make a definite etiological diagnosis. Here, we designed a custom probe library which contains the known pathogenic genes reported to be associated with Leukoencephalopathies, and performed targeted gene capture and massively parallel sequencing (MPS) among 49 Chinese patients who has white matter damage as the main imaging changes, and made the validation by Sanger sequencing for the probands’ parents. As result, a total of 40.8% (20/49) of the patients identified pathogenic mutations, including four associated with metachromatic leukodystrophy, three associated with vanishing white matter leukoencephalopathy, three associated with mitochondrial complex I deficiency, one associated with Globoid cell leukodystrophy (or Krabbe diseases), three associated with megalencephalic leukoencephalopathy with subcortical cysts, two associated with Pelizaeus-Merzbacher disease, two associated with X-linked adrenoleukodystrophy, one associated with Zellweger syndrome and one associated with Alexander disease. Targeted capture and MPS enables to identify mutations of all classes causing leukoencephalopathy. Our study combines targeted capture and MPS technology with clinical and genetic diagnosis and highlights its usefulness for rapid and comprehensive genetic testing in the clinical setting. This method will also expand our knowledge of the genetic and clinical spectra of leukoencephalopathy.

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confidence: 99%

The use of targeted genomic capture and massively parallel sequencing in diagnosis of Chinese Leukoencephalopathies

Wang

Yin

et al. 2016

Sci Rep

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“…Short tandem repeat (STR) markers were detected for potential maternity contamination analysis and identity testing with an identification detection kit (R1004T; GENESKY, Shanghai, China) according to the instruction manuals. The whole genomes of lymphocytes of the families or single blastomeres of embryos were amplified with a Qiagen whole genome amplification kit (150345#REPLI-g Single Cell Kit (96), QIAGEN/A) and sequenced by karyomapping ( Natesan et al, 2014 ) or by targeted NGS technique, an array-based gene chip described previously ( Chen et al, 2016 ). Based on informative SNPs, which is the same location in the parents but homozygous in one and heterozygous in the other, co-segregating with the identified mutation, haplotype analysis was carried out to confirm the carrier status in the family and the inheritance of embryos.…”

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confidence: 99%

Case Report: Preimplantation Genetic Testing and Pregnancy Outcomes in Women With Alport Syndrome

Shi

Chen

et al. 2021

Front. Genet.

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BackgroundAlport syndrome, a monogenic kidney disease, is characterized by progressive hemorrhagic nephritis, sensorineural hearing loss, and ocular abnormalities. Mutations in COL4A5 at Xq22 accounts for 80–85% of X-linked Alport syndrome patients. Three couples were referred to our reproductive genetics clinic for prenatal or preconception counseling.MethodsPrenatal diagnoses were performed by amplifying targeted regions of COL4A5. Targeted next-generation sequencing (NGS)-based haplotype analysis or karyomapping was performed in two patients. Pregnancy outcomes in the three patients were collected and analyzed. Published Alport syndrome cases were searched in Pubmed and Embase.ResultsPrenatal diagnoses in two cases showed one fetus harbored the same pathogenic mutation as the proband and the other was healthy. The couple with an affected fetus and the patient with a family history of Alport syndrome chose to take the preimplantation genetic testing (PGT) procedure. One unaffected embryo was transferred to the uterus, and a singleton pregnancy was achieved, respectively. Two patients presented non-nephrotic range proteinuria (<3 g/24 h) during pregnancy and the three cases all delivered at full-term. However, published Alport cases with chronic kidney disease or proteinuria during pregnancy were came with a high rate (75%) of adverse maternal and fetal outcomes.ConclusionThe PGT procedure performed in this study was proven to be practicable and might be expanded to be applied in other monogenic diseases. Moderate or severe renal impairments in Alport syndrome were strongly associated with adverse maternal and fetal outcomes, and baseline proteinuria was a potential predictor for pregnancy outcomes of Alport syndrome as other kidney diseases.

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“…Since the first application of PGT in 1989 [13], it has been used for various single-gene disorders and for hereditary cancer predisposition syndromes [4, 14]. At present, >150 types of single-gene disorders can be detected through PGT-M [15]. The thousands of apparently healthy offspring suggest that PGT-M is a safe and reliable procedure without marked adverse effects.…”

Section: Introductionmentioning

confidence: 99%

Preimplantation Genetic Testing Prevented Intergenerational Transmission of X-Linked Alport Syndrome

Zhang

Yuan

et al. 2021

Kidney Dis

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Background: Alport syndrome (AS) is a hereditary renal basement membrane disease that can lead to end-stage renal disease in young adults. It can be diagnosed by genetic analysis, being mostly caused by mutations in COL4A3, COL4A4, and COL4A5. To date, there is no radical cure for this disease. Objectives: The aim of this study was to avoid the transmission of AS within an affected family by selecting healthy embryos for uterine transfer. The embryos were identified by preimplantation genetic testing for monogenic disorders (PGT-M). Methods: We used next-generation sequencing (NGS) to identify mutations in the proband and his parents. The results of NGS were confirmed by Sanger sequencing. Targeted NGS combined with targeted single-nucleotide polymorphism haplotyping was used for the in vitro identification of COL4A5 mutations in human embryos to prevent their intergenerational transmission. Results: The c.349_359delGGACCTCAAGG and c.360_361insTGC mutations in COL4A5 were identified in a family affected by X-linked AS. Whole-genome sequencing by NGS with targeted haplotyping was performed on biopsied trophectoderm cells. A healthy baby was born after transfer of a single freeze-thawed blastocyst. Conclusions: The use of targeted NGS for identifying diagnostic markers combined with targeted haplotyping is an easy and efficient PGT-M method for preventing intergenerational transmission of AS.

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Identification of PKD2 mutations in human preimplantation embryos in vitro using a combination of targeted next-generation sequencing and targeted haplotyping

Cited by 15 publications

References 12 publications

The use of targeted genomic capture and massively parallel sequencing in diagnosis of Chinese Leukoencephalopathies

The use of targeted genomic capture and massively parallel sequencing in diagnosis of Chinese Leukoencephalopathies

Case Report: Preimplantation Genetic Testing and Pregnancy Outcomes in Women With Alport Syndrome

Preimplantation Genetic Testing Prevented Intergenerational Transmission of X-Linked Alport Syndrome

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